Process for removing bile salts from a patient and compositions therefor

ABSTRACT

A method for removing bile salts from a patient by ion exchange by administering to the patient a therapeutically effective amount of one or more highly crosslinked polymers characterized by a repeat unit having the formula ##STR1## or copolymer thereof, where n is an integer; R 1  is H or a C 1  -C 8  alkyl group: M is ##STR2## Z is O, NR 3 , S, or (CH 2 ) m  ; m=0-10; R 3  is H or a C 1  -C 8  alkyl group; and R 2  is ##STR3## where p=0-10, and each R 4 , R 5 , and R 6 , independently is H, a C 1  -C 8  alkyl group, or an aryl group.

RELATED APPLICATION

This application is a continuation-in-part of U.S. Ser. No. 08/258,477filed Jun. 10, 1994, which is a continuation-in-part of Ser. No.08/071,564, filed Jun. 2, 1993 and now abandoned. This application isalso a continuation-in-part of U.S. Ser. No. 08/258,431, filed Jun. 10,1994. This application also relates to U.S. Ser. No. 08/332,096, filedOct. 31, 1994, and U.S. Ser. No. 08/460,980, entitled "Process forRemoving Bile Salts from a Patient and Alkylated Compositions Therefor,"filed Jun. 5, 1995, and U.S. Ser. No. 08/471,769, entitled "AminePolymer Sequestrant and Method of Cholesterol Depletion," filed Jun. 6,1995. Each of the above applications are incorporated by reference intheir entirety.

BACKGROUND OF THE INVENTION

Salts of bile acids act as detergents to solubilize and consequently aidin digestion of dietary fats. Bile acids are precursors to bile salts,and are derived from cholesterol. Following digestion, bile acids can bepassively absorbed in the jejunum, or, in the case of conjugated primarybile acids, reabsorbed by active transport in the ileum. Bile acidswhich are not reabsorbed are deconjugated and dehydroxylated bybacterial action in the distal ileum and large intestine.

Reabsorption of bile acids from the intestine conserves lipoproteincholesterol in the bloodstream. Conversely, blood cholesterol level canbe diminished by reducing reabsorption of bile acids.

One method of reducing the amount of bile acids that are reabsorbed isoral administration of compounds that sequester the bile acids andcannot themselves be absorbed. The sequestered bile acids consequentlyare excreted.

Many bile acid sequestrants, however, do not bind conjugated primarybile acids, such as conjugated choleic and chenodeoxycholic acids wellenough to prevent substantial portions from being reabsorbed. Inaddition, the volume of sequestrants that can be ingested safely islimited. As a result, the effectiveness of sequestrants to diminishblood cholesterol levels is also limited.

A need exists, therefore, for a sequestrant and a method which overcomesor minimizes the referenced problems.

SUMMARY OF THE INVENTION

In a first aspect, the invention features a method of removing bilesalts from a patient by ion exchange that includes administering to thepatient a therapeutically effective amount of one or more crosslinkedpolymers that are essentially non-toxic and stable once ingested. Thepolymers are characterized by a repeat unit having the formula ##STR4##or copolymer thereof, where n is an integer; R¹ is H or an alkyl group(which may be straight chain or branched, substituted or unsubstituted,e.g., a C₁ -C₈ alkyl, such as methyl); M is ##STR5## or --Z--R² ; Z isO, NR³, S, or (CH₂)_(m) ; m=0-10; R³ is H or an alkyl group (which maybe straight chain or branched, substituted or unsubstituted, e.g., C₁-C₈ alkyl, such as methyl) ;and R² is ##STR6## where p=0-10, and eachR⁴, R⁵, and R⁶, independently, is H, an alkyl group (which may bestraight chain or branched, substituted or unsubstituted, e.g., C₁ -C₈alkyl, such as methyl), or an aryl group (e.g., having one or more ringsand which may be substituted or unsubstituted, e.g., phenyl, naphthyl,imidazolyl, or pryridyl).

In preferred embodiments, the polymer is crosslinked by means of amultifunctional crosslinking co-monomer, the co-monomer being present inan amount from about 0.5-25% (more preferably about 2.5-20% (or about1-10%)) by weight, based upon total monomer weight.

The invention provides an effective treatment for removing bile saltsfrom a patient (and thereby reducing the patient's cholesterol level).The compositions are non-toxic and stable when ingested intherapeutically effective amounts.

The invention further provides an effective synthesis for polymershaving hydrophilic and hydrophobic units by conducting the reaction inthe presence of an alcoholic solvent not normally considered a goodpolymerization solvent due to its chain transfer properties.

DETAILED DESCRIPTION OF THE INVENTION

As set forth, the invention relates to polymers and their use insequestering bile salts. In preferred embodiments, the polymer ischaracterized by the formula: ##STR7## where R¹ is hydrogen or methylwhere hereinafter n is an integer, unless otherwise specified, Z¹ is Oor NR³, R³ is hydrogen or an alkyl group, R⁴, R⁵ and R⁶ are,independently, hydrogen or methyl, and p=2-10.

In another preferred embodiment, the polymer is characterized by theformula: ##STR8## wherein R¹ is hydrogen or methyl, R⁴, R⁵ and R⁶ are,independently hydrogen or alkyl and p=0-2.

The polymer can also be characterized by the formula ##STR9## wherein R¹is hydrogen or methyl where hereinafter m=0-10, unless otherwisespecified, R³ is hydrogen or an alkyl group, R⁴, R⁵ and R⁶ are,independently, hydrogen or methyl, and p=2-10.

The polymers also include heteropolymers of two or more of the above.

The polymer further can include one or more hydrophobic co-monomers,e.g., styrene, vinyl naphthalene, ethyl vinylbenzene, N-alkyl and N-arylderivatives of acrylamide and methacrylamide, alkyl and aryl acrylates,alkyl and aryl methacrylates, 4-vinylbiphenyl, 4-vinylanisole,4-aminostyrene, and fluorinated derivatives of any of these co-monomers(e.g., p-fluorostyrene, pentafluorostyrene, hexafluoroisopropylacrylate,hexafluorobutylmethacrylate, or heptadecafluoro-decylmethacrylate). Forexample, the hydrophobic co-monomer can be an alkylated derivative ofone or more of the above mentioned formula. The alkyl groups arepreferably C₁ -C₁₅ (e.g., C₁ -C₁₅ alkyl groups, and may be straightchain, branched, or cyclic (e.g., cyclohexyl), and may further besubstituted or unsubstituted. The aryl groups preferably have one ormore rings and may be substituted or unsubstituted, e.g., phenyl,naphthyl, imidazolyl, or pyridyl. The polymer may also include one ormore positively charged or amine co-monomers, e.g., vinyl pyridine,dimethylaminomethyl styrene, or vinyl imidazole.

One example of a preferred polymer is characterized by a repeat unithaving the formula ##STR10## or copolymer thereof. R¹ is hydrogen ormethyl and p=2-6. The polymer may further include, as a co-monomer, oneor more of the following: n-butylmethacrylamide,hexafluorobutylmethacrylate, heptadecafluorodecylmethacrylate, styreneor fluorinated derivatives thereof, 2-vinyl naphthalene, 4-vinylimidazole, vinyl pyridine, trimethylammoniumethylmethacrylate,trimethylammoniumethylacrylate, 4-vinylbiphenyl, 4-vinylanisole, or4-aminostyrene.

Examples of suitable fluorinated styrene derivatives includep-fluorostyrene and pentafluorostyrene. Examples of suitable fluorinatedalkyl methacrylates include hexafluorobutyl methacrylate andheptadecafluorodecyl methacrylate.

A second example of a preferred polymer is characterized by a repeatunit having the formula ##STR11## or copolymer thereof. Where p=2-6. Thepolymer may also include, as a co-monomer, one or more of the following:isopropylacrylamide, styrene or fluorinated derivatives thereof,hexafluoroisopropylacrylate, and trimethylammoniumethylmethacrylate.

A third example of a preferred polymer is characterized by a repeat unithaving the formula ##STR12## or copolymer thereof wherein P is asdefined above. The polymer may also include, as a co-monomer, styrene ora fluorinated derivative thereof.

A fourth example of a preferred polymer is characterized by a repeatunit having the formula ##STR13## or copolymer thereof. Wherein p=0-10.

A fifth example of a preferred polymer is characterized by a repeat unithaving the formula ##STR14## or copolymer thereof. Wherein p=0-10.

A sixth example of a preferred polymer is characterized by a repeat unithaving the formula ##STR15## or copolymer thereof. The polymer mayfurther include, as a co-monomer, ethyl vinylbenzene.

A seventh example of a preferred polymer is characterized by a repeatunit having the formula ##STR16## or copolymer thereof.

An eighth example of a preferred polymer is characterized by a repeatunit having the formula ##STR17## or copolymer thereof wherein p=0-10.The polymer may also include, as a co-monomer, styrene or a fluorinatedderivative thereof.

In another aspect, the invention features polymers and a method forremoving bile salts from a patient by ion exchange that includesadministering to the patient a therapeutically effective amount of oneor more crosslinked polymers characterized by a repeat unit having theformula ##STR18## or copolymer thereof, where n is an integer; R¹ is Hor a C₁ -C₈ alkyl group; L is --NH-- or ##STR19## G is ##STR20## andeach R², R³, and R⁴, independently, is H, a C₁ -C₈ alkyl group, or anaryl group. The polymers are preferably non-toxic and stable onceingested.

In preferred embodiments, the polymer is crosslinked by means of amultifunctional crosslinking co-monomer which is present in an amountfrom about 0.5-25% by weight (and preferably from about 2.5-20% (e.g.about 1-10%) by weight), based upon total monomer weight. The polymerfurther can include one or more of the above-described hydrophobicco-monomers. Advantageously, cross-linking the polymer renders thepolymer non-digestible and non-absorbable or "stable" in the patient.

By "stable" it is meant that when ingested in therapeutically effectiveamounts the polymers do not dissolve or otherwise decompose to formpotentially harmful by-products, and remain substantially intact so thatthey can transport ions following ion exchange out of the body.

The polymers are preferably crosslinked by adding a crosslinkingco-monomer to the reaction mixture during polymerization. Examples ofsuitable crosslinking co-monomers include diacrylates anddimethacrylates (e.g., ethylene glycol diacrylate, propylene glycoldiacrylate, butylene glycol diacrylate, ethylene glycol dimethacrylate,propylene glycol dimethacrylate, butylene glycol dimethacrylate,polyethyleneglycol dimethacrylate, polyethyleneglycol diacrylate),methylene bisacrylamide, methylene bismethacrylamide, ethylenebisacrylamide, ethylenebismethacrylamide, ethylidene bisacrylamide,divinyl benzene, bisphenol A dimethacrylate, and bisphenol A diacrylate.These crosslinking co-monomers are either commercially available or areprepared as described in Mandeville et al., "Process for Adjusting IonConcentration in a Patient and Compositions Therefor," U.S. Ser. No.08/065,113, filed May. 20, 1993, assigned to the same assignee as thepresent application and hereby incorporated by reference. The amount ofcrosslinking agent is typically between 1.0 and 25 weight %, based uponcombined weight of crosslinking agent and monomer, with 2.5-20% beingpreferred.

One example of a preferred polymer is characterized by a repeat unithaving the formula ##STR21## or copolymer thereof. The polymer mayfurther include, as a co-monomer, styrene or a fluorinated derivativethereof.

A second example of a preferred polymer is characterized by a repeatunit having the formula ##STR22## or copolymer thereof.

Preferably, the polymer includes one or more co-monomers that increasethe overall hydrophobicity of the polymer. Because bile salts arehydrophobic, the hydrophobic co-monomer aids in maximizing theselectivity of the interaction of the polymer with the bile salts.

Examples of suitable hydrophobic co-monomers include, e.g., acrylamide,methacrylamide, and N-alkyl (e.g., methyl, ethyl, isopropyl, butyl,hexyl, dodecyl, cyclohexyl, dicyclohexyl) and N-aryl (e.g., phenyl,diphenyl) derivatives thereof; alkyl and aryl acrylates andmethacrylates (e.g., ethyl, propyl, butyl, dodecyl), and fluorinatedderivatives thereof (e.g., hexafluoroisopropyl acrylate, hexafluorobutylmethacrylate, heptadecafluorodecyl acrylate); styrene and derivativesthereof (e.g., dimethylaminomethyl styrene, 4-aminostyrene, andfluorinated derivatives, e.g., p-fluorostyrene, pentafluorostyrene);ethylvinylbenzene; vinyl napthalene; vinyl pyridine; vinyl imidazole;4-vinylbiphenyl; 4,4-vinylanisole; and combinations thereof. The amountof hydrophobic co-monomer used in the preparation of these polymers isfrom 1 to 75% by weight, preferably from 3 to 65%.

The level of hydrophobicity needed may also be achieved simply byappropriate choice of crosslinking co-monomer. For example,divinylbenzene is a suitable crosslinking co-monomer and is hydrophobicas well. In addition, the main "impurity" in divinylbenzene isethylvinylbenzene, a hydrophobic, polymerizable monomer which will alsocontribute to the overall hydrophobicity of the polymer. Otherhydrophobic crosslinking co-monomers include bisphenol A diacrylate andbisphenol A dimethacrylate.

The polymers can have fixed positive charges, or may have the capabilityof becoming charged upon ingestion at physiological pH. In the lattercase, the charged ions also pick up negatively charged counterions uponingestion that can be exchanged with bile salts. In the case of polymershaving fixed positive charges, however, the polymer may be provided withone or more exchangeable counterions, as a pharmaceutically acceptablesalt.

By "salt" it is meant that the amine nitrogen group in the repeat unitis protonated to create a positively charged nitrogen atom associatedwith a negatively charged counterion.

Examples of suitable counterions include Cl⁻, Br⁻, CH₃ OSO₃ ⁻, HSO₄ ⁻,SO₄ ², HCO₃ ⁻, CO₃ ² ⁻, acetate, lactate, succinate, propionate,butyrate, ascorbate, citrate, maleate, folate, an amino acid derivative,a nucleotide, a lipid, or a phospholipid. The counterions may be thesame as, or different from, each other. For example, the polymer maycontain two different types of counterions, both of which are exchangedfor the bile salts being removed. More than one polymer, each havingdifferent counterions associated with the fixed charges, may beadministered as well.

By "non-toxic" it is meant that when ingested in therapeuticallyeffective amounts neither the polymers nor any ions released into thebody upon ion exchange are substantially harmful. Preferably, the ionsreleased into the body are actually beneficial to the patient. Such isthe case when, for example, the exchangeable ions are natural nutrientssuch as amino acids, or possess a therapeutic value.

As set forth above, preferred polymers of the claimed invention areco-polymers of the amine or ammonium monomeric units of the aboveformulae and one or more hydrophobic monomers. The co-polymers can beprepared by co-polymerization of the monomers. Alternatively oradditionally, one or more of the amine or ammonium monomeric units canbe rendered hydrophobic through alkylation.

Thus, the present invention includes reaction products of (a) one ormore cross-linked polymers characterized by repeat unit of the formula:##STR23## wherein R¹, M and n are as defined above; and (b) at least onealkylating agent.

Each of the preferred polymers described above can be subjected to thealkylation reaction.

For example, the polymer can be the reaction product of:

(a) one or more crosslinked polymers characterized by a repeat unithaving the formula: ##STR24## and salts and copolymers thereof, where nand m are integers, and each R¹, R², and R³, independently, is H or a C₁-C₈ alkyl group; and

(b) at least one alkylating agent. The reaction product is preferablynon-toxic and stable once ingested.

Cross-linking can be achieved during or subsequent to the polymerizationof polymer (a). Alternatively, the reaction product of (a) and (b) canbe crosslinked by means of a multifunctional crosslinking co-monomer,the co-monomer being present in an amount from about 0.5-25% (morepreferably about 2.5-20% (e.g., about 1-10%)) by weight, based on totalweight monomer weight.

By "alkylating agent" it is meant a reactant which, when reacted withthe crosslinked polymer, causes an alkyl group or derivative thereof(e.g., substituted alkyls, such as an aralkyl, hydroxyalkyl,alkylammonium salt, alkylamide, or combination thereof) to be covalentlybound to one or more of the nitrogen atoms of the polymer.

Preferred alkylating agents have the formula RX where R is a C₁ -C₂₀alkyl, C₁ -C₂₀ hydroxyalkyl, C₁ -C₂₀ aralkyl, C₁ -C₂₀ alkylammonium, orC₁ -C₂₀ alkylamido group and X includes one or more electrophilicleaving groups. By "electrophilic leaving group" it is meant a groupwhich is displaced by a nitrogen atom in the crosslinked polymer duringthe alkylation reaction. Examples of preferred leaving groups includehalide, epoxy, tosylate, and mesylate group. In the case of, e.g., epoxygroups, the alkylation reaction causes opening of the three-memberedepoxy ring.

Examples of preferred alkylating agents include a C₁ -C₂₀ alkyl halide(e.g., C₄ -C₁₂ alkyl halide) an-butyl halide, n-hexyl halide, n-octylhalide, n-decyl halide, n-dodecyl halide, n-tetradecyl halide,n-octadecyl halide, and combinations thereof; a C₁ -C₂₀ dihaloalkane(e.g., a 1,10-dihalodecane); a C₁ -C₂₀ hydroxyalkyl halide (e.g., an11-halo-1-undecanol); a C₁ -C₂₀ aralkyl halide (e.g., a benzyl halide);a C₁ -C₂₀ alkyl halide ammonium salt (e.g., a(4-halobutyl)trimethylammonium salt, (6-halohexyl)trimethylammoniumsalt, (8-halooctyl)trimethylammonium salt,(10-halodecyl)trimethylammonium salt, (12-halododecyl)trimethylammoniumsalts and combinations thereof); a C₁ -C₂₀ alkyl epoxy ammonium salt(e.g., a (glycidylpropyl)trimethylammonium salt); and a C₁ -C₂₀ epoxyalkylamide (e.g., an N-(2,3-epxypropane)butyramide,N-(2,3-epoxypropane)hexanamide, and combinations thereof).

It is particularly preferred to react the polymer with at least twoalkylating agents. In one preferred example, one of the alkylatingagents has the formula RX where R is a C₁ -C₂₀ alkyl group and Xincludes one or more electrophilic leaving groups (e.g., an alkylhalide), and the other alkylating agent has the formula R'X where R' isa C.sub. -C₂₀ alkyl ammonium group and X includes one or moreelectrophilic leaving groups (e.g., an alkyl halide ammonium salt).

In another preferred example, one of the alkylating agents has theformula R'X where R is a C₁ -C₂₀ alkyl group and X includes one or moreelectrophilic leaving groups (e.g., an alkyl halide), and the otheralkylating agent has the formula R'X where R' is a C₁ -C₂₀ hydroxyalkylgroup and X includes one or more electrophilic leaving groups (e.g., ahydroxy alkyl halide).

In another preferred example, one of the alkylating agents is a C₁ -C₂₀dihaloalkane and the other alkylating agent is a C₁ -C₂₀ alkylammoniumsalt.

EXAMPLES

A. Polymer Preparation

1. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride(PolyMAPTAC)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (40mL of a 50% aqueous solution, 21 g), ethylene glycol dimethacrylatecrosslinking co-monomer (5.00 g, 4.76 mL), ethyl acetate (200 mL), and2-propanol (200 mL). The resulting solution was clear. Next, thepolymerization initiator AIBN (0.1 g) was added and the reaction mixturewas heated to 65° C. When the temperature reached 65° C., the solutionwas degassed with nitrogen for 5 minutes, at which point it turnedcloudy, indicating that polymerization was proceeding. The reaction wasmaintained at 65° C. for another 3 hours and then allowed to cool toroom temperature.

The resulting polymer (which was hard and sticky) was combined with 500mL of water to soften it, and then transferred to a blender where it wasblended with 1500 mL of 2-propanol and centrifuged. The mixture was thendecanted and transferred to another blender with the aid of 100 mL ofwater. 800 mL of 2-propanol was then added and the mixture was blended,allowed to settle, and decanted. The mixture was then combined with 1000mL of 2-propanol, blended, filtered, and vacuum-dried to afford 12.6 gof polymer.

PolyMAPTAC crosslinked with 0.5% methylenebismethacrylamide crosslinkingco-monomer; polyMAPTAC crosslinked with 10% methylenbismethacrylamidecrosslinking co-monomer; and polyMAPTAC crosslinked with 10%divinylbenzene crosslinking co-monomer were prepared in analogousfashion.

2. Preparation of Poly(vinylamine)

The first step involved the preparation of ethylidenebisacetamide.Acetamide (118 g), acetaldehyde (44.06 g), copper acetate (0.2 g), andwater (300 mL) were placed in a 1 L three neck flask fitted withcondenser, thermometer, and mechanical stirrer. Concentrated HCl (34 mL)was added and the mixture was heated to 45°-50° C. with stirring for 24h. The water was then removed in vacuo to leave a thick sludge whichformed crystals on cooling to 5° C. Acetone (200 mL) was added andstirred for a few minutes, after which the solid was filtered off anddiscarded. The acetone was cooled to 0° C. and solid was filtered off.This solid was rinsed in 500 mL acetone and air dried 19 h to yield 31.5g of ethylidenebisacetamide.

The next step involved the preparation of vinylacetamide fromethylidenebisacetamide. Ethylidenebisacetamide (31.05 g), calciumcarbonate (2 g) and celite 541 (2 g) were placed in a 500 mL three neckflask fitted with a thermometer, a mechanical stirrer, and a distillinghead atop a Vigreux column. The mixture was vacuum distilled at 35 mm Hgby heating the pot to 180°-225° C. Only a single fraction was collected(10.8 g) which contained a large portion of acetamide in addition to theproduct (determined by NMR). This solid product was dissolved inisopropanol (30 mL) to form the crude vinylacetamide solution used forpolymerization.

Crude vinylacetamide solution (15 mL), divinylbenzene (1 g, technicalgrade, 55% pure, mixed isomers), and AIBN (0.3 g) were mixed and heatedto reflux under a nitrogen atmosphere for 90 min, forming a solidprecipitate. The solution was cooled, isopropanol (50 mL) was added, andthe solid was collected by centrifugation. The solid was rinsed twice inisopropanol, once in water, and dried in a vacuum oven to yield 0.8 g,of poly(vinylacetamide), which was used to prepare poly(vinylamine asfollows.

Poly(vinylamide) (0.79 g) was placed in a 100 mL one neck flaskcontaining water (25 mL) and conc. HCl (25 mL). The mixture was refluxedfor 5 days, after which the solid was filtered off, rinsed once inwater, twice in isopropanol, and dried in a vacuum oven to yield 0.77 gof product. Infrared spectroscopy indicated that a significant amount ofthe amide (1656 cm⁻¹) remained and that not much amine (1606 cm⁻¹) wasformed. The product of this reaction (˜0.84 g) was suspended in NaOH (46g) and water (46 g ) and heated to boiling (˜140° C.). Due to foaming,the temperature was reduced and maintained at ˜100° C. for 2 h. Water(100 mL) was added and the solid collected by filtration. After rinsingonce in water the solid was suspended in water (500 mL) and adjusted topH 5 with acetic acid. The solid was again filtered off, rinsed withwater, than isopropanol, and dried in a vacuum oven to yield 0.51 g ofproduct. Infrared spectroscopy indicated that significant amine had beenformed.

3. Preparation of Poly(3-dimethylaminopropylacrylamide) (DMAPA)

Dimethylaminopropylacrylamide (10 g) and methylenebisacrylamidecrosslinking co-monomer (1.1 g) were dissolved in 50 mL of water in a100 mL three neck flask. The solution was stirred under nitrogen for 10minutes. Potassium persulfate (0.2 g) and sodium metabisulfite (0.2 g)were each dissolved in 2-3 mL of water and then mixed. After a fewseconds this solution was added to the monomer solution, still undernitrogen. A gel formed immediately and was allowed to sit overnight. Thegel was removed and blended with 500 mL of isopropanol. The solid wasfiltered off and rinsed three times with acetone. The solid white powerwas filtered off and dried in a vacuum oven to yield 61.1 g.

4. Preparation of Poly(3-dimethylaminopropymethylamide)hydrochloride)(DMAPA.HCl)

Dimethylaminopropymethylamide (20.0 g) was dissolved in water (100 mL)and neutralized with concentrated HCl to pH 6.95. Methylenebisacrylamidecrosslinking co-monomer (2.2 g) and water (100 mL) were added and warmed(34° C.) to dissolve. Potassium persulfate (0.2 g) and potassiummetabisulfite (0.2 g) were added with stirring. After gelation, thesolution was allowed to sit for 6 h, blended with isopropanol (600 mL)three times, and dried in a vacuum oven to yield 14.47 g of the titlepolymer.

PolyDMAPA.HCl crosslinked with 10% methylenebismethacrylamidecrosslinking co-monomer was prepared in analogous fashion.

5. Preparation of Poly(3-dimethylaminopropymethacrylamide)hydrochloride)(DMAPMA.HCl)

Dimethylaminopropylmethacrylamide (20.0 g) was dissolved in water (100mL) and neutralized with concentrated HCl to pH 6.94.Methylenebisacrylamide crosslinking co-monomer (2.2 g) was added and thesolution was warmed (39° C.) to dissolve. Potassium persulfate (0.2 g)and potassium metabisulfite (0.2 g) were added with stirring under anitrogen atmosphere. After gelation, the solution was allowed to sitovernight, blended with isopropanol (500 mL) twice, and dried in avacuum oven t6 yield 27.65 g of product. Some of the solid (3.2 g;sieved to -80/+200 mesh size) was stirred in water (100 mL) for 50 min,additional water (100 mL) was added and the solution stirred for 36minutes. The solid was collected by centrifugation, resuspended in water(400 mL), stirred 150 minutes, and again collected by centrifugation.The solid was finally resuspended in water (500 mL), stirred 90 minutes,and collected by filtration. The solid was dried in a vacuum oven toyield 0.28 g of the title polymer.

6. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(n-butylmethacrylamide) (MAPTAC co-BuMA)

The co-monomer n-butylmethacrylamide (BuMA) was prepared as follows.

Methacryloyl chloride (48.4 mL, 52.3 g, 0.500 mol) was dissolved intetrahydrofuran (300 mL) in a 1 L flask and placed in an ice bath. Asolution containing butylamine (36.6 g) and triethylamine (55.6 g) wasadded dropwise, maintaining the temperature at 5°-15° C. After additionthe solution was stirred for 5 minutes and the solid triethylaminehydrochloride was filtered off and discarded. The solvent was removed invacuo from the mother liquor and the resulting yellow oil was usedwithout further purification. The yield was 71.58 g of BuMA co-monomer.

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (108mL of a 50% aqueous solution, 56.8 g), ethylene glycol dimethacrylatecrosslinking co-monomer (19.62 g), BuMA co-monomer (12.12 g), and2-propanol (850 mL). The resulting solution was clear. Next, thereaction mixture was heated to 40° C. while being degassed withnitrogen. When the solution had reached 40° C., the catalyst, consistingof a solution of potassium persulfate (0.75 g) and potassiummetabisulfate (0.75 g) in 25 mL of water was added. The solutionimmediately began to turn cloudy, indicating the polymerization wasproceeding. The reaction was maintained at 40° C. for 24 hours and thenallowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and vacuum dried to afford 64.54 g of the title polymer.

Polymer for testing was washed two times with 800 mL of water each time,followed by two washes with 500 mL of methanol each time to give 34.5 gof purified polymer.

A crosslinked MAPTAC co-BuMA copolymer was also prepared using propyleneglycol dimethacrylate, rather than ethylene glycol dimethacrylate, asthe crosslinking co-monomer, as follows.

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (60mL of a 50% aqueous solution, 31.5 g), propylene glycol dimethacrylatecrosslinking co-monomer (9.81 g), BuMA co-monomer (6.06 g), and2-propanol (300 mL). The resulting solution was clear. Next, thereaction mixture was heated to 70° C. while being degassed withnitrogen. When the solution had reached 70° C., the catalyst, AIBN (0.50g), was added. The solution immediately began to turn cloudy, indicatingthat polymerization was proceeding. The reaction was maintained at 70°C. for 6 hours and then allowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and vacuum dried to afford 23.3 g of polymer.

MAPTAC coBuMA (5%) crosslinked with 24% ethyleneglycoldimethacrylatecrosslinking co-monomer, MAPTAC coBuMA (20%) crosslinked with 0.5%methylenebismethacrylamide crosslinking co-monomer, and MAPTAC coBuMA(14%) crosslinked with 22% propyleneglycoldimethacrylate crosslinkingco-monomer were prepared in analogous fashion by adjusting the ratios ofstarting monomers.

7. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(styrene) (MAPTAC co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (60mL of a 50% aqueous solution, 31.5 g), divinyl benzene crosslinkingco-monomer (2.00 styrene co-monomer (1.75 g), and 2-propanol (300 mL).The resulting solution was clear. Next, the reaction mixture was heatedto 60° C. while being degassed with nitrogen. When the solution hadreached 60° C., the catalyst, AIBN (0.50 g), was added. The solutionimmediately began to turn cloudy, indicating that polymerization wasproceeding. The reaction was maintained at 60° C. for 24 hours and thenallowed to cool to room temperature. After about 7 hours the mixture hadbecome very thick and 100 mL additional isopropanol was added to allowfor better stirring.

The resulting polymer was filtered and washed on the funnel withisopropanol and vacuum dried to afford 30.9 of the title polymer.

Polymer for testing was washed two times with 1000 mL of water each timefollowed by two washes with 800 mL of methanol each time to give 28.0 gof purified polymer.

MAPTAC co-Sty (13%) crosslinked co-monomer, MAPTAC co-Sty (13%)crosslinked with 20% butyleneglycoldimethacrylate crosslinkingco-monomer, MAPTAC co-Sty (19%) crosslinked with 6% divinylbenzeneco-monomer, MAPTAC co-Sty (23%) crosslinked with 7% divinylbenzeneco-monomer, MAPTAC co-Sty (30%) crosslinked with 6% divinylbenzeneco-monomer, and MAPTAC co-Sty (38%) crosslinked with 6% divinylbenzeneco-monomer were prepared in analogous fashion by varying the ratios ofstarting monomers.

8. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(vinyl naphthalene) (MAPTAC co-VN)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (40mL of a 50% aqueous solution, 21.0 g), divinyl benzene crosslinkingco-monomer (2.25 g), 2-vinylnaphthalene co-monomer (10.5 g), and2-propanol (320 mL). The resulting solution was clear. Next, thereaction mixture was heated to 65° C. while being degassed withnitrogen. When the solution had reached 65° C., the catalyst, AIBN (0.50g), was added. The solution immediately began to turn cloudy, indicatingthat polymerization was proceeding. The reaction was maintained at 64°C. for 20 hours and then allowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and then immediately slurried in 400 mL of distilled water.The mixture was stirred for 1/2 hour and then filtered. The water washwas repeated one more time. The filter cake was then slurried in 400 mLof methanol and stirred for 1/2 hour. The mixture was filtered and themethanol slurry was repeated one more time. Vacuum drying afforded 22.11g, 65.5% of the title polymer.

MAPTACco-VN (39%) crosslinked with 5% divinyl benzene crosslinkingco-monomer was prepared in analogous fashion by varying the ratio ofstarting monomers.

9. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(1-vinyl imidazole) MAPTAC co-VI)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC) (40mL of a 50% aqueous solution, 1.0 g), divinyl benzene crosslinkingco-monomer (2.25 g), 1-vinylimidazole co-monomer (12.54 g), and2-propanol (300 mL). The resulting solution was clear. Next, thereaction mixture was heated to 65° C. while being degassed withnitrogen. When the solution had reached 65° C., the catalyst, AIBN (0.50g), was added. The solution immediately began to turn cloudy, indicatingthat polymerization was proceeding. The reaction was maintained at 65°C. for 20 hours and then allowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol, and then immediately slurried in 500 mL of distilled water.The mixture was stirred for 1/2 hour and then filtered. The water washwas repeated one more time. The filter cake was then slurried in 400 mLof methanol and stirred for 1/2 hour. The mixture was filtered and themethanol slurry was repeated one more time. Vacuum drying afforded 7.34g, 20.5% of the title polymer.

10. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(styrene) TMAEAC co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: trimethylammoniumethylacrylatechloride (TMAEAC) (99.4 mL of a50% aqueous solution, 53.0 g), divinyl benzene crosslinking co-monomer(7.00 g), styrene co-monomer (40.0 g), and 2-propanol (800 mL). Theresulting solution was clear. Next, the reaction mixture was heated to65° C. while being degassed with nitrogen. When the solution had reached65° C., the catalyst, AIBN (1.50 g), was added. The solution immediatelybegan to turn cloudy, indicating that polymerization was proceeding. Thereaction was maintained at 65° C. for 6 hours, then cooled to 60° C. andstirred for an additional 18 hours. It was then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol, and then immediately slurried in 1000 mL of distilledwater. The mixture was stirred for 1/2 hour and then 800 mL of methanolwas added and the mixture was stirred for an additional 1/2 hour. Themixture was allowed to settle and the supernatant liquid was decanted,leaving a residue of about 750 mL. The residue was then slurried with anadditional 750 mL of methanol and stirred for 1/2 hour. The methanolslurry and decantation process was repeated two more times with 800 mLof methanol each time. Next, 800 mL of isopropanol was added and themixture was stirred for 1/2 hour and then filtered. Finally, 600 mL ofisopropanol was added and the mixture was stirred for 1/2 hour.Filtration and vacuum drying afforded 49.2 g, 49.2% of the titlepolymer.

TMAEAC co-Sty (31%) crosslinked with 8% divinylbenzene crosslinkingco-monomer and TMAEAC co-Sty (46%) crosslinked with 6% divinylbenzenecrosslinking co-monomer were prepared in analogous fashion by varyingthe ratio of starting monomers.

11. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(styrene) TMAEAC co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: Trimethylammoniumethylmetharylate chloride (TMAEMAC) (38.0 mLof a 50% aqueous solution, 21.7 g), divinyl benzene crosslinkingco-monomer (3.72 g), styrene co-monomer (15.66 g), and 2-propanol (2500mL). The resulting solution was clear. Next, the reaction mixture washeated to 65° C. while being degassed with nitrogen. When the solutionhad reached 65° C., the catalyst, AIBN (0.50 g), was added. The solutionimmediately began to turn cloudy, indicating that polymerization wasproceeding. After two hours, the mixture became very thick and anadditional 100 mL of isopropanol was added. After five hours the mixturewas again very thick so an additional 100 mL of isopropanol was added.The reaction was maintained at 65° C. for 6 hours, and then allowed tocool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol, and then immediately slurried in 1000 mL of distilledwater. The mixture was stirred for 1/2 hour and then transferred to ablender and blended for five minutes. The polymer slurry was filteredand 1000 mL of distilled water was added and the mixture was stirred for1/2 hour. The mixture was filtered and the filter cake was slurried twotimes in 500 mL of methanol each time. Filtration and vacuum dryingafforded 30.2 g, 75.9% of the title polymer.

TMAEMC co-Sty (58%) crosslinked with 4% divinylbenzene crosslinkingco-monomer, TMAEMC co-Sty (33%) crosslinked with 4% divinylbenzenecrosslinking co-monomer, and TMAEMC co-Sty (24%) crosslinked with 4%divinylbenzene crosslinking co-monomer were prepared in analogousfashion by varying the ratio of starting the monomers.

12. Preparation ofPoly(methacrylamidopropyl-3-(trimethylammonium)chloride) co-(poly2,3,4,5,6-pentafluorostyrene) (MAPTAC co-StyF₅)

To a 1000 mL, three-necked, round-bottom flask was added the following:methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC) (24.5 mL ofa 50% aqueous solution, 13.00 g), divinylbenzene crosslinking co-monomer(1.00 g), pentafluorostyrene (6.00 g), 2-propanol (150 mL), and AIBN(0.50 g). The resulting solution was clear. Next, the reaction mixturewas heated to 65° C. while being degassed with nitrogen. After a shortperiod of time, the solution began to turn cloudy, indicating thatpolymerization was proceeding. After five hours the mixture was verythick so an additional 100 mL of isopropanol was added. The reaction wasmaintained at 65° C. for 24 hours, and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of distilled water. Themixture was stirred for 1/2 hour. The polymer slurry was filtered and500 mL of distilled water was added and the mixture was stirred for 1/2hour. The mixture was filtered and the filter cake was slurried twotimes in 300 mL of methanol each time. Filtration and air dryingafforded 7.74 g of the title co-polymer.

MAPTAC co-StyF₅ (20%) crosslinked with 5% divinylbenzene crosslinkingco-monomer, MAPTAC co-StyF₅ (40%) crosslinked with 5% divinylbenzenecrosslinking co-monomer, and MAPTAC co-StyF₅ (45%) crosslinked with 5%divinylbenzene crosslinking co-monomer were prepared in analogousfashion by varying the ratio of starting monomers.

13. Preparation ofpoly(methacrylamidopropyl-3-(trimethylammonium)chloride), co-poly2-(trimethylammonium)ethyl methacrylate chloride, co-styrene (MAPTACco-TMAEMC co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC)(10.40 g of a 50% aqueous solution, 5.20 g), 2-(trimethylammonium)ethylmethacrylate chloride (TMAEMC) (4.86 g of a 70% aqueous solution, 3.40g) divinylbenzene crosslinking co-monomer (1.00 g), styrene (10.40 g),2-propanol (150 mL), and AIBN (0.50 g). The resulting solution wasclear. Next, the reaction mixture was heated to 70° C. while beingdegassed with nitrogen. After a short period of time, the solution beganto turn cloudy, indicating that polymerization was proceeding. Thereaction was maintained at 70° C. for 24 hours, and then allowed to coolto room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of methanol. The mixturewas stirred for 1/2 hour. The polymer slurry was filtered and 400 mL ofdistilled water was added and the mixture was stirred for 1/2 hour. Themixture was filtered and the water slurry was repeated. The mixture wasfiltered and the filter cake was slurried two times in 400 mL ofmethanol each time. Filtration and air drying afforded 5.39 g of thetitle co-polymer.

MAPTAC co-TMAEMC (34%) co-Sty (36%) crosslinked with 5% divinylbenzenecrosslinking co-monomer, MAPTAC co-TMAEMC (31%) co-Sty (41%) crosslinkedwith 5% divinylbenzene crosslinking co-monomer, MAPTAC co-TMAEMC (28%)co-Sty (46%) crosslinked with 5% divinylbenzene crosslinking co-monomer,MAPTAC co-TMAEMC (23%) co-Sty (48%) crosslinked with 5% divinylbenzenecrosslinking co-monomer, MAPTAC co-TMAEMC (26%) co-Sty (52%) crosslinkedwith 4% divinylbenzene crosslinking co-monomer, MAPTAC co-TMAEMC (15%)co-Sty (55%) crosslinked with 4% divinylbenzene crosslinking co-monomer,and MAPTAC co-TMAEMC (13%) co-Sty (61.5%) crosslinked with 4%divinylbenzene crosslinking co-monomer were prepared in analogousfashion by varying the ratio of starting monomers.

14 Preparation of Poly(trimethylammoniumethylmethacrylatechlorideco-poly(isopropylacrylamide)) (TMAEMAC co-IPA)

The co-monomer isopropylacrylamide (IPA) was first prepared as follows:

Acryloyl chloride (63 mL, 70.2 g, 0.775 mol) was dissolved intetrahydrofuran (200 mL) in a 1 L flask and placed in an ice bath. Asolution containing isopropylamine (127.7 mL, 88.67 g, 1.50 mol) wasadded dropwise, maintaining the temperature at 5-15° C. After additionthe solution was stirred for 10 minutes and the solid isopropylaminehydrochloride was filtered off and discarded. The solvent was removed invacuo from the mother liquor and the resulting almost colorless oil,which solidified on standing, was used without further purification toprepare the title co-polymer as follows.

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: trimethylammoniumethylacrylate chloride (76.5 mL of a 50%aqueous solution, 41.18 g, 0.213 mol), methylene bis acrylamidecrosslinking co-monomer (2.40 g), IPA co-monomer (4.52 g, 0.070 mol),and water (200 mL). The resulting solution was clear. The reactionmixture was stirred while being degassed with nitrogen. When thesolution had been degassed, the catalyst, consisting of potassiumpersulfate (0.3 g) and potassium metabisulfate (0.2 g) was added. Thepolymerization initiated after 2 minutes and gelled after 3 minutes.

The next morning the gel was transferred to a blender and 1000 mL ofwater was added. After blending for a few seconds, the polymer hadswelled to take up all of the water. The swollen polymer was blended inseveral portions with isopropanol several times to dehydrate it. Theresulting polymer was filtered and washed on the funnel with isopropanoland vacuum dried to afford 36.8 g of the title co-polymer.

15. Preparation of Poly(methacrylamidopropyl-3-(trimethylammoniumchloride) co-poly(vinyl pyridine) (MAPTAC co-VP)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC)(40 mL of a 50% aqueous solution, 21.0 g), divinyl benzene crosslinkingco-monomer (2.25 g), vinyl pyridine (14.0 g, 0.133 mol), conc.hydrochloric acid (11 mL, 0,133 mol), 2-propanol (300 mL), and AIBN(0.67 g). The resulting solution was clear. Next, the reaction mixturewas heated to 60° C. for 20 hours, and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 1000 mL of distilled water. Themixture was stirred for 1 hour. The polymer slurry was filtered, washedon the funnel with methanol, and then slurried in 600 mL of methanol forone hour. Filtration and air drying afforded 20.4 g of co-polymer.

16. Preparation of Poly(trimethylammoniumethylmethacrylate chloride)co-poly(p-fluorostyrene) (TMAEMC co-F₁ Sty)

To a 500 mL flask was added the following:trimethylammoniumethylmethacrylate chloride (TMAEMC) (11.0 g of a 70%aqueous solution, 7.70 g), divinylbenzene crosslinking co-monomer (0.50g), p-fluorostyrene co-monomer (4.00 g), 2-propanol (125 mL) and AIBN(0.25 g). The resulting solution was clear. Next, the reaction mixturewas heated to 65° C. while being degassed with nitrogen. The solutionimmediately began to turn cloudy, indicating that polymerization wasproceeding. The reaction was maintained at 65° C. for 6 hours, and thenallowed to cool to room temperature.

The solvent was removed by decantation and the polymer was immediatelyslurried in 250 mL of distilled water. The mixture was stirred for 1/2hour and then decanted. The water slurry was repeated three more times.Finally, the polymer was slurried in 400 mL of methanol. Filtration andvacuum drying afforded 5.42 g, 44.4% of the title co-polymer.

TMAEMC co-F₁ Sty (24%) crosslinked with 4% divinylbenzene crosslinkingco-monomer was prepared in analogous fashion by varying the ratio of thestarting monomers.

17. Preparation of Poly(methacrylamidopropyl-3-(trimethyl ammoniumchloride)) co-poly(hexafluorobutyl methacrylate) (MAPTAC coF₆ BMA)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium) chloride (MAPTAC)(28.5 mL of a 50% aqueous solution, 15.0 g), divinylbenzene crosslinkingco-monomer (1.00 g), hexafluorobutyl methacrylate (4.00 g), 2-propanol(150 mL), and AIBN (0.50 g). The resulting solution was clear. Next, thereaction mixture was heated to 60° C. while being degassed withnitrogen. After a short period of time, the solution began to turncloudy, indicating that polymerization was proceeding. The reaction wasmaintained at 60° C. for 24 hours, and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of distilled water. Themixture was stirred for 1 hour. The polymer slurry was filtered and thewater slurry was repeated one more time. The polymer was then slurriedin 500 mL of methanol for one hour and filtered. The methanol slurry wasrepeated one more time. Finally, the polymer was slurried in 400 mL ofisopropanol and stirred overnight. Filtration and air drying afforded7.52 g of the title co-polymer.

18. Preparation of Poly(trimethylammoniumethylacrylate chloride)co-poly(hexafluoroisopropyl acrylate) (TMAEAC co-F₆ IA )

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: trimethylammoniumethylacrylate chloride (30.0 mL of a 50%aqueous solution, 15.0 divinylbenzene crosslinking co-monomer (1.00 g),F₆ IPA co-monomer (4.00 g), AIBN (0.50 g), and isopropanol (150 Theresulting solution was clear. The reaction mixture was stirred whilebeing degassed with nitrogen and heated to 60° C. After 18 hours thereaction mixture was allowed to cool to room temperature and the solventwas removed by decanting. The residual polymer was slurried in 400 mL ofwater, stirred for one hour and filtered. The water slurry was repeatedone more time. Next, the polymer was slurried two times in methanol.Finally, the polymer was slurried in 200 mL of isopropanol, stirred fortwo hours and filtered. Air drying afforded 5.59 g of the title polymer.

19. Preparation of Poly(methacrylamidopropyl-3-(trimethylammoniumchloride)) co-poly(heptadecafluorodecyl methacrylate) (MAPTAC coF₁₇DecMA)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium chloride (MAPTAC)(28.5 mL of a 50% aqueous solution, 15.0 g), divinylbenzene crosslinkingco-monomer (1.00 g), heptadecafluorodecyl methacrylate (4.00 g),2-propanol (150 mL), and AIBN (0.40 g). The result solution was clear.Next, the reaction mixture was heated to 65° C. while being degassedwith nitrogen. After a short period of time, the solution began to turncloudy, indicating that polymerization was proceeding. After four hours,the reaction mixture had gotten very thick and 100 mL more isopropanolwas added. The reaction was maintained at 65° C. for 18 hours, and thenallowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 600 mL of distilled water. Themixture was stirred for 1 hour. The polymer slurry was filtered and thewater slurry was repeated one more time. The polymer was then slurriedin 500 mL of methanol for one hour and filtered. Air drying afforded17.73 g of co-polymer.

20. Preparation of poly(methacrylamidopropyl-3-(trimethylammoniumchloride)), co-poly(2-(trimethylammonium)ethyl acrylate chloride),co-poly styrene (MAPTAC co-TMAEAC co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC)(10.00 g of a 50% aqueous solution, 5.00 g), 2-(trimethylammonium)ethylmethacrylate chloride (TMAEAC) (6.00 g of a 50% aqueous solution, 3.00g) divinylbenzene crosslinking co-monomer (1.00 styrene (11.00 g),2-propanol (150 mL), and the AIBN (0.25 g). The resulting solution wasclear. Next, the reaction mixture was heated to 70° C. while beingdegassed with nitrogen. After a short period of time, the solution beganto turn cloudy, indicating that polymerization was proceeding. Thereaction was maintained at 70° C. for 24 hours, and then allowed to coolto room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of methanol. The mixturewas stirred for 1/2 hour. The polymer slurry was allowed to settle anddecanted. 200 mL of distilled water was added and the mixture wasstirred for 1/2 hour. The mixture was decanted and the water slurry wasrepeated with 400 mL. The mixture was decanted and the polymer wasslurried two times in 200 mL of methanol each time. Filtration and airdrying afforded 2.76 g of the title co-polymer.

MAPTAC co-TMAEAC (10%) co-Sty (60%) crosslinked with 5% divinylbenzenecrosslinking co-monomer was prepared in analogous fashion by varying theratio of starting monomers.

21. Preparation of poly(2-(trimethylammonium)ethyl acrylate chloride)co-poly(2,3,4,5,6-pentafluorostyrene) (TMAEAC co-StyF₅)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: 2-(trimethylammonium)ethyl acrylate chloride (TMAEAC) (24.0mL of a 50% aqueous solution, 13.00 g), divinylbenzene crosslinkingco-monomer (1.00 pentafluorostyrene (6.00 g), 2-propanol (150 mL), andthe AIBN (0.50g). The resulting solution was clear. Next, the reactionmixture was heated to 65° C. while being degassed with nitrogen. After ashort period of time, the solution began to turn cloudy, indicating thatpolymerization was proceeding. After two hours the mixture was verythick so an additional 100 mL of isopropanol was added. The reaction wasmaintained at 65° C. for 22 hours, and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 400 mL of distilled water. Themixture was stirred for 1/2 hour. The polymer slurry was filtered and600 mL of distilled water was added and the mixture was stirred for 1/2hour. The mixture was filtered and the filter cake was slurried in 400mL of methanol. Filtration and air drying afforded 7.26 g of the titleco-polymer.

TMAEAC co-StyF₅ (20%) crosslinked with 5% divinylbenzene crosslinkingco-monomer was prepared in analogous fashion by varying the ratio ofstarting monomers.

22. Preparation of poly(2-(trimethylammonium)ethyl methacrylatechloride), co-poly(2,3,4,5,6-pentafluorostyrene) (TMAEMC co-StyF₅)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: 2-(trimethylammonium)ethyl methacrylate chloride (TMAEMC)(19.52 of a 70% aqueous solution, 13.66 g), divinylbenzene crosslinkingco-monomer (1.00 g), pentafluorostyrene (9.18 g), 2-propanol (150 mL),and AIBN (0.40 g). The resulting solution was clear. Next, the reactionmixture was heated to 70° C. while being degassed with nitrogen. After ashort period of time, the solution began to turn cloudy, indicating thatpolymerization was proceeding. After 1.5 hours the mixture was verythick so an additional 50 mL of isopropanol was added. The reaction wasmaintained at 70° C. for 5 hours, and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of distilled water. Themixture was stirred for 1/4 hour. The polymer slurry was filtered and500 mL of distilled water was added and the mixture was stirred for 1/4hour. The water slurry was repeated one more time. The mixture wasfiltered and the filter cake was slurried three times in 300 mL ofmethanol each time. Filtration and air drying afforded 1.26 g of thetitle co-polymer.

TMAEMC co-StyF₅ (24%) crosslinked with 4% divinylbenzene crosslinkingco-monomer and TMAEMC co-StyF₅ (39%) crosslinked with 4% divinylbenzenecrosslinking co-monomer were prepared in analogous fashion by varyingthe ratio of starting monomers.

23. Preparation of Poly(ethyleneimine)

Polyethyleneimine (120 g of a 50% aqueous solution; Scientific PolymerProducts) was dissolved in water (250 mL). Epichlorohydrin (22.1 mL) wasadded dropwise. The solution was heated to 60° C. for 4 h, after whichit had gelled. The gel was removed, blended with water (1.5 L) and thesolid was filtered off, rinsed three times with water (3 L) and twicewith isopropanol (3 L), and the resulting gel was dried in a vacuum ovento yield 81.2 g of the title polymer.

24. Preparation of Poly(methacrylamidopropyl-3-(trimethylammoniumchloride)), co-poly(2-(trimethylammonium)ethylmethacrylate chloride))co-poly(2,3,4,5,6-pentafluorostyrene) (MAPTAC co-TMAEMC co-StyF₅)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC)(10.00 of a 50% aqueous solution, 5.00 g), 2-(trimethylammonium)ethylmethacrylate chloride (TMAEMC) (5.71 g of a 70% aqueous solution, 4.00g), divinylbenzene crosslinking co-monomer (1.00 g), pentafluorostyrene(10.00 g), 2-propanol (150 mL), and AIBN (0.50 g). The resultingsolution was clear. Next, the reaction mixture was heated to 70° C.while being degassed with nitrogen. After a short period of time, thesolution began to turn cloudy, indicating that polymerization wasproceeding. The reaction was maintained at 70° C. for 24 hours, and thenallowed to cool to room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 500 mL of methanol. The mixturewas stirred for 1/4 hour. The of methanol. The mixture was stirred for1/4 hour. The polymer slurry was filtered and then slurried 3 times in300 mL of water each time. The last time the polymer slurry was blendedfor 5 minutes. The mixture was filtered and the filter cake was slurriedtwo times in 300 mL of methanol each time. Filtration and vacuum dryingafforded 9.74 g of co-polymer.

25. Preparation of poly(2-(trimethylammonium)ethyl acrylate chloride),co-poly(2-(trimethylammonium)ethylmethacrylate chloride) co-styrene(TMAEAC, co-TMAEMC, co-Sty)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: 2-(trimethylammonium)ethyl acrylate chloride (TMAEAC) (6.00 gof a 50% aqueous solution, 3.00 g), 2-(trimethylammonium)ethylmethacrylate chloride (TMAEMC) (4.29 g of a 70% aqueous solution, 3.00g), divinylbenzene crosslinking co-monomer (1.00 g), styrene (13.00 g),2-propanol (150 mL), and AIBN (0.50 g). The resulting solution wasclear. Next, the reaction mixture was heated to 70° C. while beingdegassed with nitrogen. After a short period of time, the solution beganto turn cloudy, indicating that polymerization was proceeding.

The reaction was maintained at 70° C. for 24 hours, and then allowed tocool to room temperature.

The resulting polymer was decanted and immediately slurried in 500 mL ofmethanol. The mixture was stirred for 1/2 hour. The polymer slurry wasfiltered and 500 mL of distilled water was added. The mixture was thenstirred for 1/2 hour and blended for 10 minutes. The mixture was allowedto settle and the water was decanted. The water slurry was repeated twomore times and the decantation residue was slurried two times in 400 mLof methanol each time, settling and decanting each time. Vacuum dryingafforded 8.03 g of the title co-polymer.

26. Preparation ofPoly(methacrylamidopropyl-3-(trimethylammonium)chloride),co-poly(2-(trimethylammonium)ethylacrylate chloride)co-poly(2,3,4,5,6-pentafluorostyrene) (MAPTAC co-TMAEAC co-StyF₅)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyl-3-(trimethylammonium)chloride (MAPTAC)(8.00 g of a 50% aqueous solution, 4.00 g), 2-(trimethylammonium)ethylacrylate chloride (TMAEMA) (6.00 g of a 50% aqueous solution, 3.00 g),divinylbenzene crosslinking co-monomer (1.00 pentafluorostyrene (12.00g), 2-propanol (150 mL), and AIBN (0.50g). The resulting solution wasclear. Next, the reaction mixture was heated to a 70° C. while beingdegassed with nitrogen. After a short period of time, the solution beganto turn cloudy, indicating that polymerization was proceeding. Thereaction was maintained at 70° C. for 24 hours, and then allowed to coolto room temperature.

The resulting polymer was filtered and washed on the funnel withisopropanol and immediately slurried in 400 mL of methanol. The mixturewas stirred for 1/2 hour. The polymer slurry was filtered and thenslurried 2 times in 250 mL of water each time. The last time the polymerslurry was blended for 5 minutes. The mixture was filtered and thefilter cake was slurried two times in 250 mL of methanol each time.Filtration and vacuum drying afforded 7.80 g of co-polymer.

27. Preparation of Poly((trimethylammonium)ethyl acrylate chloride),co-poly(2-(trimethylammonium)ethyl methacrylate chloride)co-poly(2,3,4,5,6-pentafluorostyrene) (TMAEAC, co-TMAEMC, co-StyF₅)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: 2-(trimethylammonium)ethyl acrylate chloride (TMAEAC) (61.00g of a 50% aqueous solution, 3.00 g), 2-(trimethylammonium)ethylmethacrylate chloride (TMAEMC) (4.29 g of a 70% aqueous solution, 3.00g), divinylbenzene crosslinking co-monomer (1.00 g), pentafluorostyrene(13.00 g), 2-propanol (150 mL), and AIBN (0.50g). The resulting solutionwas clear. Next, the reaction mixture was heated to 70° C. while beingdegassed with nitrogen. After a short period of time, the solution beganto turn cloudy, indicating that polymerization was proceeding. Thereaction was maintained at 70° C. for 24 hours, and then allowed to coolto room temperature.

The resulting polymer was decanted and immediately slurried in 400 mL ofmethanol. The mixture was stirred for 1/2 hour. The polymer slurry wasfiltered and then slurried two times in 200 mL of water each time. Thesecond time the polymer slurry was blended for 5 minutes. The mixturewas filtered and the filter cake was slurried two times in 200 mL ofmethanol each time. Vacuum drying afforded 6.87 g of co-polymer.

28. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(4-vinylbiphenyl) (MAPTAC co-VBPh)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC)(10.49 g of a 50% aqueous solution, 0.0475 mol), 4-vinylbiphenyl (VBPh)(9.01 g, 0.050 mol), divinyl benzene crosslinking co-monomer (1.47 g),2-propanol (150 mL), and the polymerization initiator AIBN (0.25 g). Theresulting mixture contained insoluble VBPh which dissolved upon warming.Next, the reaction mixture was heated to 70° C. while degassing withnitrogen. After a short period of time, the solution began to turncloudy, indicating that polymerization was proceeding. The reaction wasmaintained at 70° C. for 24 hours and then filtered while hot.

The resulting polymer was washed on the filtration funnel withisopropanol, and then immediately slurried in 200 mL of methanol,followed by stirring for 1 hour. The polymer slurry was then filtered,after which the methanol slurry procedure was repeated. Next, thepolymer was slurried two times using 200 mL of water each time. Theresulting mixture was then filtered and the filter cake slurried twotimes with methanol using 200 mL of methanol each time. The resultingmixture was then filtered and vacuum dried to afford 9.59 g ofco-polymer.

29. Preparation of Poly(methacrylamidopropyltrimethylammonium chloride)co-poly(4-vinylanisole) (MAPTAC co-VA)

To a 1000 mL, three-necked, round-bottomed flask was added thefollowing: methacrylamidopropyltrimethylammonium chloride (MAPTAC)(10.49 g of a 50% aqueous solution, 0.0475 mol), 4-vinylanisole (VA)(6.71 g, 0.050 mol), divinyl benzene crosslinking co-monomer (1.30 g),2-propanol (200 mL), and the polymerization initiator AIBN (0.40 g). Theresulting clear solution was heated to 70° C. while degassing withnitrogen. After several hours the solution began to turn cloudy,indicating that polymerization was proceeding. The reaction wasmaintained at 70° C. for 36 hours and then allowed to cool to roomtemperature.

The resulting polymer was filtered and washed on the filtration funnelwith isopropanol, and then immediately slurried in 200 mL of methanol,followed by stirring for 1 hour. The polymer slurry was then filteredand slurried two times using 200 mL of water each time. The resultingmixture was then filtered and the filter cake slurried in 200 mL ofmethanol, after which it was slurried in 200 mL of isopropanol. Theresulting mixture was then filtered and vacuum dried to afford 5.19 g ofco-polymer.

30. Preparation ofPoly(N-(4-methylstyrene)-N'-(3-trimethylammonio-2-hydroxypropylchloride)piperazine)

The first step in the reaction is the preparation of4-(piperazinylmethyl)styrene).

To a 500 mL flask was added vinylbenzyl chloride (7.63 g, 0.050 mol),piperazine (8.61 g, 0.100 mol), and isopropanol (50 mL). The resultingsolution was heated to 70° C. for 45 minutes and then cooled slowly toroom temperature to form a slurry of crystalline material. The slurrywas refrigerated for about three hours and then filtered. The solidpiperazine hydrochloride salt was vacuum dried and then weighed (5.55 g)and discarded. The mother liquor was concentrated to about 25 mL on arotary evaporator and ethyl acetate (50 mL) was added). The resultingmixture was refrigerated for about 10 minutes, after which a second cropof piperazine hydrochloride was filtered off and discarded. The motherliquor was then evaporated to dryness on a rotary evaporator to afford10.135 g of crude 4-(piperazinylmethyl)styrene which was used withoutfurther purification to prepareN-(4-methylstyrene)-N'-(3-trimethylammonio-2-hydroxypropylchloride)piperazine as follows.

To a 500 mL flask was added 4-(piperazinylmethyl)styrene (10.35 g, about0.45 mol), glycidyl trimethyl ammonium chloride (7.58 g, 0.045 mol), andisopropanol (50 mL). The resulting mixture was heated to 60° C. andstirred for 20 hours, after which it was cooled to room temperature andused for polymerization reactions without further purification asfollows.

To 0.40 mol ofN-(4-methylstyrene)-N'-(3-trimethylammonio-2-hydroxypropylchloride)piperazine in 100 mL of isopropanol was added divinyl benzenecrosslinking co-monomer (0.93 g). The resulting solution was degassedwith nitrogen and then the polymerization initiator AIBN (0.20 g) wasadded. The temperature was raised to 70° C. with continued nitrogendegassing and held there for three hours, at which point a largequantity of cross-linked polymer had precipitated. The mixture was thencooled to 40° C. and filtered.

The resulting polymer was washed on the filtration funnel withisopropanol and then immediately slurried in 200 mL of methanol, afterwhich it was stirred for 1 hour. The polymer slurry was then filteredand the methanol slurry procedure repeated. The resulting polymer wasthen slurried two times in 200 mL of water each time, followed byfiltration. The filter cake was slurried two times in 200 mL of methanoleach time. Filtration and vacuum drying afforded 7.90 g of polymer.

31. Preparation ofPoly(N-(4-methylstyrene)-N'-(3-trimethylammonio-2-hydroxypropylchloride)piperazine),co-polystyrene

To 0.022 mol ofN-(4-methylstyrene)-N'-(3-trimethylammonio-2-hydroxypropyl)piperazine(prepared as described in Example 30) was added styrene co-monomer (2.29g, 0.022 mol) and divinyl benzene crosslinking co-monomer (0.50 g, 0.004mol). The resulting solution was degrassed with nitrogen, after whichthe polymerization initiator AIBN (0.3 g) was added. The temperature wasraised to 70° C. with continued nitrogen degassing and held there forfour hours, at which point a large quantity of crosslinked polymer hasprecipitated. Twenty five mL's of additional isopropanol was then addedand heating continued for another 19 hours, after which the mixture wascooled to room temperature and filtered to yield solid polymer.

The resulting polymer was filtered and washed on the filtration funnelwith methanol and then immediately slurried in 200 mL of methanol. Themixture was stirred for 0.5 hour, after which it was filtered and thepolymer slurried two times using 250 mL of water each time. Next, themixture was filtered and the filter cake was slurried in 200 mL ofmethanol, filtered, and then slurried in 200 mL of isopropanol.Filtration and vacuum drying afforded 4.98 g of copolymer.

A copolymer containing a 2:1 molar ratio of styrene to quaternary aminemonomer was also prepared in an analogous manner.

32. Preparation ofPoly(N-(3-trimethylammonio-2-hydroxypropylchloride)-4-aminostyrene)

The first step is the preparation of 4-aminomethylstyrene as follows.

To a 250 mL flask was added vinylbenzyl chloride (7.63 g, 0.050 mol),concentrated aqueous ammonia (9.8 mL), and isopropanol (40 mL). Themixture was stirred for one week, at which point a large quantity ofcrystalline material (ammonium chloride) had precipitated. The solid wasfiltered off and washed with isopropanol. The mother liquor wasevaporated on a rotary evaporator until no ammonia odor could bedetected. Isopropanol (50 mL) was then added, and the mixturerefrigerated for several hours. Following refrigeration, ammoniumchloride which had precipitated was filtered off to yield4-aminomethylstyrene which was used without further purification.

To a 500 mL flask was added 4-aminomethylstyrene (0.050 mol), glycidyltrimethylammonium chloride (7.62 g, 0.050 mol), water (about 2 mL) toeffect solution, and divinylbenzene crosslinking co-monomer (0.98 g).The resulting solution was heated at 70° C. for 5 hours, after which asmall amount of water (about 5-10 mL) was added to dissolve someprecipitated salts. The polymerization initiator AIBN (0.20 g) was addedwhile concurrently degassing the solution with nitrogen. The reactionmixture became very thick with polymer, requiring the addition ofisopropanol to permit stirring.

After stirring for about 16 hours at 70° C., the mixture was cooled toroom temperature and filtered. The resulting polymer was washed on thefiltration funnel with methanol and then immediately slurried in 200 mLof methanol. The resulting mixture was stirred for 1 hour, after whichthe polymer slurry was filtered and the methanol slurry procedurerepeated. The polymer was then slurried two times in 200 mL of watereach time, after which the mixture was filtered and the filter cakeslurried in 200 mL of methanol. Filtration and vacuum drying afforded8.68 g of polymer.

33. Alkylation of Poly(dimethylaminopropylmethacrylamide) crosslinkedwith methylenebismethacrylamide with 1-iodooctane alkylating agent

Poly(dimethylaminopropylmethacrylamide) crosslinked withmethylenebismethacrylamide prepared as described in Example 5 (1.0 g)was suspended in methanol (100 mL) and sodium hydroxide (0.2 g) wasadded. After stirring for 15 minutes, 1-iodooctane (1.92 mL) was addedand the mixture stirred at 60° C for 20 hours. The mixture was thencooled and the solid filtered off. Next, the solid was washed bysuspending it in isopropanol (500 mL), after which it was stirred for 1hour and then collected by filtration. The wash procedure was thenrepeated twice using aqueous sodium chloride (500 mL of a 1M solution),twice with water (500 mL), and once with isopropanol (500 mL) beforedrying in a vacuum oven at 50° C.) for 24 hours to yield 0.1 g ofalkylated product.

34. Alkylation of Poly(dimethylaminopropylacrylamide) crosslinked withmethylenebismethacrylamide with 1-iodooctane alkylating agent

Poly(dimethylaminopropylacrylamide) crosslinked withmethylenebismethacrylamide prepared as described in Example 4 (10 g) wasalkylated according to the procedure described in Example 33. Theprocedure yielded 2.95 g of alkylated product.

35. Preparation of Poly 92-(methacroylamido)ethyltrimethylammoniumiodide) co-polystyrene

The first step is the preparation of 2-(N',N'-dimethylamino)-N-ethylmethacrylamide hydrochloride as follows.

To a 1000 mL flask was added methacryloyl chloride (52.3 g, 0.5 mol) andtetrahydrofuran (300 mL). The solution was cooled to less than 10° C.,after which a solution of N,N-dimethylaminoethylamine (30.5 g, 0.35 mol)in tetrahydrofuran (100 mL) was added dropwise while maintaining thetemperature at 8°-10° C. When the addition was complete, the mixture wasfiltered, washed with cold tetrahydrofuran, and vacuum dried to yield65.69 g of 2-(N',N'-dimethylamino)-N-ethyl methacrylamide hydrochloride.

The next step was the preparation of2-(methacryloylamido)ethyltrimethylammonium iodide as follows.

Potassium hydroxide (15.4 g, 0.24 mol) and methanol (240 mL) were addedto a 500 mL flask, and the mixture stirred to effect completedissolution of the potassium hydroxide. To the solution was added2-(N',N'-dimethylamino)-N-ethyl methacrylamide hydrochloride (46.35g,0.24 mol) and the resulting mixture stirred for 0.5 hour. The mixturewas then filtered to remove potassium chloride and the filtrate wasconcentrated on a rotary evaporator. Isopropanol (400 mL) and methyliodide (18.7 mL, 42.6 g, 0.30 mol) were added to the concentratedfiltrate and the mixture stirred at room temperature overnight. In themorning, the solid product was filtered off, washed with isopropanol,and vacuum dried to yield 61.08 g of2-(methacryloylamido)ethyltrimethylammonium iodide as a whitecrystalline solid.

Next, 2-(methacryloylamido)ethyltrimethylammonium iodide (12.24 g, 0.050mol), styrene (5.2 g, 0.050 mol),divinylbenzene crosslinking co-monomer(0.65 g, 0.005 mol), isopropanol (150 mL), water (20 mL), and thepolymerization initiator AIBN (0.4 g) were added to a 1000 mL flask andthe resulting solution degassed with nitrogen while heating to 70° C.The solution was then stirred for 24 hours at 70° C. under nitrogen,after which it was cooled to room temperature. At this point, thesolvent was decanted and 200 mL of methanol added to the flask to createa slurry which was stirred overnight. The product was then filtered andadded to a blender with 500 mL of water. The resulting mixture wasblended for 15 minutes and then filtered. The remaining solid materialwas washed sequentially with water (200 mL) and methanol (200 mL).Filtration and vacuum drying yielded 3.22 g of the title polymer.

Testing of Polymers

A. Preparation of Artificial Intestinal Fluid Test procedure No. 1

Sodium carbonate (1.27 g) and sodium chloride (1.87 g) were dissolved in400 mL of distilled water. To this solution was added a mixture ofpurified bile acids, consisting of taurocholic acid (0.138 g, 0.24mmol), glycocholic acid (0.292 g, 0.60 mmol), glycodeoxycholic acid(0.085 mmol, 0.18 mmol) and glycochenodeoxycholic acid (0.085 mmol, 0.18mmol). The pH of the solution was adjusted to 7.20 with acetic acid.This solution was used for the testing of the various polymers. Thetotal bile salt concentration in this solution is 3 millimolar, aconcentration approximately equal to that found in normal physiologicalsolutions in the duodenum.

Polymers were tested as follows.

To a 40 mL centrifuge tube was added 0.25 g of polymer and 20 mL of theartificial small intestinal fluid prepared as described above. Themixture was stirred in a water bath maintained at 37° C. for threehours. The mixture was then centrifuged and the supernatant liquid,being slightly cloudy, was filtered. The filtrate was analyzed for total3-hydroxy steroid content by an enzymatic assay using 3a-hydroxy steroiddehydrogenase, as described below.

Test Procedure No. 2

Sodium carbonate (1.27 g) and sodium chloride (1.87 g) were dissolved in400 mL of distilled water. To this solution was added either glycocholicacid (1.95 g, 4.0 mmol) or glycochenodeoxycholic acid 1.89 g, 4.0 mmol)to make a 10 mM solution. The pH of the solution was adjusted to 6.8with acetic acid. These stock solutions were used for the testing of thevarious polymers.

Polymers were tested as follows.

To a 14 mL centrifuge tube was added 10 mg of polymer and 10 mL of abile salt solution in concentrations ranging from 0.1-10 mM preparedfrom 10 mM stock solution (prepared as described above) and bufferwithout bile salt in the appropriate amount. The mixture was stirred ina water bath maintained at 37° C. for three hours. The mixture was thenfiltered. The filtrate was analyzed for total 3-hydroxy steroid contentby an enzymatic assay using 3a-hydroxy steroid dehydrogenase, asdescribed below.

Enzymatic Assay for Total Bile Salt Content

Four stock solutions were prepared.

Solution 1--Tris-HCl buffer, containing 0.133M Tris, 0.666 mM EDTA at pH9.5.

Solution 2--Hydrazine hydrate solution, containing 1M hydrazine hydrateat pH 9.5.

Solution 3--NAD+ solution, containing 7 mM NAD+ at pH 7.0.

Solution 4--HSD solution, containing 2 units/mL in Tris-HCl buffer (0.03M Tris, 1 mM EDTA) at pH 7.2.

To a 3 mL cuvette was added 1.5 mL of solution 1, 1.0 mL of solution 2,0.3 mL of solution 3, 0.1 mL of solution 4 and 0.1 mL ofsupernatant/filtrate from a polymer test as described above. Thesolution was placed in a UV-VIs spectrophotometer and the absorbance(O.D.) of NADH at 340 nm was measured. The bile salt concentration wasdetermined from a calibration curve prepared from dilutions of theartificial intestinal fluid prepared as described above.

All of the polymers previously described were tested in one or both ofthe above tests and all were efficacious in removing bile salts from theartificial intestinal fluid.

Use

The polymers according to the invention may be administered orally to apatient in a dosage of about 1 mg/kg/day to about 10 g/kg/day; theparticular dosage will depend on the individual patient (e.g., thepatient's weight and the extent of bile salt removal required). Thepolymer may be administrated either in hydrated or dehydrated form, andmay be flavored or added to a food or drink, if desired to enhancepatient acceptability. Additional ingredients such as other bile acidsequestrants, drugs for treating hypercholesterolemia, atherosclerosisor other related indications, or inert ingredients, such as artificialcoloring agents may be added as well.

Examples of suitable forms for administration include pills, tablets,capsules, and powders (e.g., for sprinkling on food). The pill, tablet,capsule, or powder can be coated with a substance capable of protectingthe composition from the gastric acid in the patient's stomach for aperiod of time sufficient to allow the composition to passundisintegrated into the patient's small intestine. The polymer may beadministered alone or in combination with a pharmaceutically acceptablecarrier substance, e.g., magnesium carbonate, lactose, or a phospholipidwith which the polymer can form a micelle.

EQUIVALENTS

Those skilled in the art will know, or be able to ascertain, using nomore than routine experimentation, many equivalents to the specificembodiments of the invention described herein. These and all otherequivalents are intended to be encompassed by the following claims.

What is claimed is:
 1. A method of synthesizing a crosslinked polymerhaving hydrophilic and hydrophobic units comprising: polymerizing amixture comprising a hydrophilic vinylic monomer, a monoreactivehydrophobic vinylic co-monomer and a multifunctional crosslinkingco-monomer in an alcoholic solvent under conditions suitable for freeradical polymerization.
 2. The claim 1 wherein the hydrophilic monomerhas a formula ##STR25## where R¹ is H or a C₁ -C₈ alkyl group; M is##STR26## or --Z-- R² ; Z is O, NR³, S, or (CH₂)_(m) ; m=0-10; R³ is H--or a C₁ -C₈ alkyl group; and R² is ##STR27## where p=0-10, and each R⁴,R⁵, and R⁶, independently, is H, a C₁ -C₈ alkyl group, or an group. 3.The method of claim 2 wherein said monoreactive hydrophobic co-monomeris selected from the group consisting of styrene and fluorinatedderivatives thereof; vinyl naphthalene and fluorinated derivativesthereof; ethyl vinylbenzene and fluorinated derivatives thereof; N-alkyland N-aryl derivatives of acrylamide and methacrylamide, and fluorinatedderivatives thereof; alkyl and aryl acrylates and fluorinatedderivatives thereof; alkyl and aryl methacrylates and fluorinatedderivatives thereof; 4-vinylbiphenyl and fluorinated derivativesthereof; 4-vinylanisole and fluorinated derivatives thereof; and4-aminostyrene and fluorinated derivatives thereof.
 4. The method ofclaim 2 wherein said multifunctional crosslinking co-monomer is presentin an amount from about 0.5-25% by weight, based upon total monomerweight.
 5. The method of claim 4 wherein said crosslinking co-monomer ispresent in an amount from about 1-10% by weight, based upon totalmonomer weight.
 6. The method of claim 4 wherein said crosslinkingco-monomer is selected from the group consisting of ethyleneglycoldimethylacrylate, methylene bis methacrylamide, propyleneglycoldimethyl acrylate or divinyl benzene.
 7. The method of claim 2further comprising one or more additional amine co-monomers.
 8. Themethod of claim 7 wherein said amine co-monomer is vinyl pyridine,dimethylaminomethyl styrene, or vinyl imidazole.
 9. The method of claim2 wherein said hydrophilic monomer has a formula ##STR28## thereofwherein R¹ and p are as defined in claim
 2. 10. The method of claim 9wherein said monoreactive hydrophobic co-monomer isn-butylmethacrylamide, hexafluorobutylmethacrylate,heptadecafluorodecylmethacrylate, styrene or a fluorinated derivativethereof, 2-vinyl naphthalene, 4-vinyl imidazole, vinyl pyridine,4-vinylbiphenyl or a fluorinated derivative thereof, 4-vinylanisole or afluorinated derivative thereof, or 4-aminostyrene or a fluorinatedderivative thereof.
 11. The method of claim 9 further comprising ahydrophilic monomer selected from the group consisting oftrimethylammoniumethylmethacrylate and trimethylammoniumethylacrylate.12. The method of claim 2 wherein said hydrophilic monomer has a formula##STR29## wherein R¹ and p are as defined in claim
 2. 13. The method ofclaim 12 wherein said monoreactive hydrophobic co-monomer isisopropylacrylamide, styrene or a fluorinated derivative thereof orhexafluoroisopropylacrylate.
 14. The method of claim 12 furthercomprising the hydrophilic monomer trimethylammoniumethylmethacrylate.15. The method of claim 2 wherein said hydrophilic monomer has a formula##STR30## wherein Z is NR³ or (CH₂)_(m), R² is ##STR31## R¹, R³, R⁴, R⁵,R⁶, m and p are as defined in claim
 2. 16. The method of claim 15wherein said monoreactive hydrophobic co-monomer is selected from thegroup consisting of styrene and fluorinated derivatives thereof; vinylnaphthalene and fluorinated derivatives thereof; ethyl vinylbenzene andfluorinated derivatives thereof; N-alkyl and N-aryl derivatives ofacrylamide and methacrylamide, and fluorinated derivatives thereof;alkyl and aryl acrylates and fluorinated derivatives thereof; alkyl andaryl methacrylates and fluorinated derivatives thereof; 4-vinylbiphenyland fluorinated derivatives thereof; 4-vinylanisole and fluorinatedderivatives thereof; and 4-aminostyrene and fluorinated derivativesthereof.
 17. A method of synthesizing a crosslinked polymer havinghydrophilic and hydrophobic units comprising: polymerizing a mixturecomprising a hydrophilic vinylic monomer, a monoreactive hydrophobicvinylic co-monomer and a multifunctional crosslinking co-monomer in analcoholic solvent, under conditions suitable for free radicalpolymerization wherein said hydrophilic vinylic monomer has a repeatunit having the formula ##STR32## where R¹ is H or a C₁ -C₈ alkyl group;L is --NH-- or ##STR33## G is ##STR34## and each R², R³, independently,is H, a C₁ -C₈ alkyl group, or an aryl group.
 18. The method of claim 17wherein said multifunctional crosslinking co-monomer is present in anamount from about 0.5-25% by weight, based upon total monomer weight.19. The method of claim 18 wherein said crosslinking co-monomer ispresent in an amount from about 1-10% by weight, based upon totalmonomer weight.
 20. The method of claim 17 wherein said monoreactivehydrophobic monomer is selected from the group consisting of styrene andfluorinated derivatives thereof; vinyl naphthalene and fluorinatedderivatives thereof; ethyl vinylbenzene and fluorinated derivativesthereof; N-alkyl and N-aryl derivatives of acrylamide andmethacrylamide, and fluorinated derivatives thereof; alkyl and arylacrylates and fluorinated derivatives thereof; alkyl and arylmethacrylates and fluorinated derivatives thereof; 4-vinylbiphenyl andfluorinated derivatives thereof; 4-vinylanisole and fluorinatedderivatives thereof; and 4-aminostyrene and fluorinated derivativesthereof.
 21. The method of claim 17 wherein said hydrophilic monomer hasa formula ##STR35## wherein G is as defined in claim
 17. 22. The methodof claim 17 wherein said hydrophilic monomer has a formula ##STR36##wherein G is as defined in claim
 17. 23. A method of synthesizing acrosslinked polymer having hydrophilic and hydrophobic units comprising:polymerizing a mixture comprising hydrophilic vinylic monomers,monoreactive hydrophobic vinylic co-monomers and a multifunctionalcrosslinking co-monomer in an alcoholic solvent, under conditionssuitable for free radical polymerization wherein the alcoholic solventis 2-propanol.